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Cytoskeleton, Inc. 1830 S. Acoma St., Actin Tools Denver, CO 80223, USA.

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Tubulin Tools • Tubulin Polymerization Assays • Tubulin Binding Assays • Labeled and Unlabeled Tubulin Proteins

Live Cell Imaging Tools • Spirochrome: Actin, DNA, and Tubulin Tools • New MemGlow™ Flourogenic Probes • New Flipper Probes and SPY-BG Dyes

Activation Assay Tools • Measure Active Small G-Proteins • RhoA, cdc42, Rac1, Arf, and Ras • GLISA and Pulldown Applications

Trusted Actin Tools by Cytoskeleton, Inc. Actin Tools Actin Tools

™ Acti-stain™ 488 Acti-stain™ 535 Acti-stain Fluorescent Phalloidins phalloidin Rhodamine phalloidin and Spirochrome™ Live Cell Probes Labeled Actins Highly pure, biologically active actins labeled with Rhodamine, Pyrene, or Biotin. The Acti-stain™ line of fluorescent phalloidins has been developed with Labeled Actin Proteins an emphasis on creating exceptionally bright and stable probes for Labeled Actins Source Purity Cat. # Amount F-actin offered at an economical price. Side-by-side comparisons with Rabbit skeletal AB07-A 5 x 20 µg Biotinylated Actin Protein >99% similar products insure considerable savings without sacrificing quality muscle AB07-C 20 x 20 µg Rabbit skeletal AP05-A 1 x 1 mg Acti-stain™ 555 Acti-stain™ 670 Pyrene Actin Protein >99% when switching to an Acti-stain™ probe. The combination of in-house phalloidin phalloidin muscle AP05-B 5 x 1 mg Bovine cardiac manufacturing, stringent quality control, and convenient packaging Pyrene Actin Protein >99% CS-AP07 1 x 250 µg muscle provides a great value. Give them a try and see for yourself. Human platelet, APHR-A 4 x 10 µg Rhodamine Actin Protein >99% non-muscle APHR-C 20 x 10 µg Rabbit skeletal AR05-B 10 x 20 µg Rhodamine Actin Protein >99% For more information, citations and comparison to other muscle AR05-C 20 x 20 µg fluorescent phalloidins, visit: cytoskeleton.com/actin/acti-stain

Product Excitation Emission Signal stability without Cat. # Amount** Cytoskeleton’s Biochem Kits™ are comprehensive kits for assaying different antifade* (T1/2 in secs) aspects of Actin biochemistry! Acti-stain™ 488 phalloidin 480 nm 535 nm 57 PHDG1-A 300 Slides Acti-stain™ 535 phalloidin (Rhodamine phalloidin) 535 nm 585 nm 27 PHDR1 300 Slides Actin bundling assay using kit BK001 Acti-stain™ 555 phalloidin 535 nm 585 nm 46 PHDH1-A 300 Slides Actin Binding Protein Spin- Bundled F-actin was pelleted by a 14,000 x g -actinin α centrifugation and pellets (P) and supernatants (S) Acti-stain™ 670 phalloidin 640 nm 670 nm 18 PHDN1-A 300 Slides Down Assay Biochem Kit were run on a SDS-PAGE gel. Only in the presence of SiR700-Actin Kit 690 nm 720 nm na*** CY-SC013 35 nmol F-actin + F-actin only F-actin + BSA the F-actin bundling protein a-actinin is actin pelleted Includes SiR-Actin and Verapamil • Identifies and characterizes Actin Binding S P S P S P at this centrifugation speed. SiR-Actin Kit Proteins (ABPs) 630 nm 680 nm na*** CY-SC001 50 nmol Includes SiR700-Actin and Verapamil • Generation of saturation binding curves Product Cat. # Amount * Stability measured with stained slides without antifade. For comparison, fluorescein • Muscle (BK001) or non-muscle (BK013) actin α-actinin 2 Actin Binding Protein Spin- phalloidin has a T1/2 of 6 secs. ** One slide equals enough phalloidin to stain a 25 mm BSA 30-100 Down Assay Biochem Kit™ BK001 High Purity & Biologically Active coverslip. *** SiR was approximately ten fold more stable than Alexa647 and as stable assays AKL99 AKL95 as atto647N (Lukinavičius, et. al.; Nature Chemistry, 5, 132–139, 2013.). SiR-Actin is a This co-sedimentation assay will help you identify Actin (skeletal muscle actin) trademarks of Spirochrome SA (Switzerland). whether your ABP is a F-actin binding protein, a Actin Binding Protein 30-100 Spin-Down Assay Biochem Kit™ BK013 F-actin severing protein, has F-actin bundling activity, Spin: 14,000 x g assays (non-muscle actin) >99% Pure or is a G-actin binding protein. (Cat. AKL99) NEW Pan-Actin Antibody Reorganization of actin after AAN02 is a mouse monoclonal antibody against actin protein. The G-actin/F-actin In Vivo antibody has been shown to recognize α-skeletal, α-cardiac, α-smooth treatment with jasplakinolide ™ Swiss 3T3 cells were treated with jasplakinolide muscle, β-cytoplasmic, γ-smooth muscle and γ- cytoplasmic actin Assay Biochem Kit (Jaspl) or left untreated (Untr) and the G-actin (G) and isotypes. • Quantitates monomeric vs polymeric actin in The highest purity actin available. Actin polymerization stimulated by F-actin (F) content was assayed using the G-actin/F- Purities greater than 99% from most Arp2/3 complex and the VCA domain cell/tissue lysates actin kit. Treatment with jasplakinolide resulted in a sources. Cited hundreds of times in of WASP measured by Pyrene Actin • Reproducible and accurate method potent accumulation of F-actin. the literature. fluorescence (Cat.# AP05). • Contains all needed reagents Unlabeled Actin Proteins Lyse cells or tissue in the F-actin stabilizing buffer, Product Cat. # Amount preserving the G-actin:F-actin ratio. Centrifuge G-actin/F-actin In Vivo 30-100 BK037 Unlabeled Actins Source Purity Cat. # Amount samples, separating supernatants (G-actin) and Assay Biochem Kit™ assays AKL99-A 4 x 250 µg Protease Inhibitor Cocktail pellets (F-actin) which are then run on a gel for PIC02 1 ml AKL99-B 2 x 1 mg (100x solution) Actin Protein Rabbit skeletal muscle >99% AKL99-C 5 x 1 mg Western blot analysis. AKL99-D 10 x 1 mg AKL99-E 20 x 1 mg AKL95-B 1 x 1 mg Actin Protein Rabbit skeletal muscle >97% AKL95-C 5 x 1 mg 8000 Actin Polymerization Assay Actin + AD99-A 1 x 1 mg Characterization of ABPs using Actin Actin Protein Bovine cardiac muscle >99% Arp2/3 + AD99-B 5 x 1 mg ™ VCA Polymerization Biochem Kit™ Smooth muscle, chicken AS99-A 1 x 1 mg Biochem Kit Actin Protein >99% Immunofluorescence images of mouse Swiss 3T3 cells stained with anti- 6000 Effects of Arp2/3 (Cat. # RP01P) and the WASP VCA gizzard AS99-B 5 x 1 mg • Utilizes fluorescent pyrene-actin Actin + VCA (Cat. # VCG03) domain on actin polymerization in APHL99-A 2 x 250 µg actin antibody. Swiss 3T3 cells were grown to 25% confluency on poly- lysine and laminin coverslips. 3T3 cells were fixed with PFA. Cells were • F-actin polymerization and depolymerization Actin + Arp2/3 vitro. Arp2/3 or the WASP VCA domain alone has Actin Protein Human platelet, non-muscle >99% APHL99-C 1 x 1 mg Actin only APHL99-E 5 x 1 mg permeabilized with methanol followed by 0.5% Triton X-100 as described • Works with multiple sources of actin 4000 little effect on the rate of actin polymerization, while RFU Pre-formed Actin AKF99-A 1 x 1 mg in the method. IF staining using 1:500 dilution of anti-actin antibody in the combination of the two leads to an activation Rabbit skeletal muscle >99% • Valuable for characterizing ABPs Filaments AKF99-B 5 x 1 mg PBS is shown (green). of the actin nucleating Arp2/3 complex and a Actin Thin Filament subsequent increased rate of actin polymerization. Bovine cardiac muscle 90% TFC01 1 x 1 mg This kit is based upon the enhanced fluorescence 2000 (Ca2+ sensitive complex) Product Cat. # Amount of pyrene-conjugated actin that occurs during Actin Thin Filament Anti-Pan Actin Mouse Monoclonal Antibody (Clone Rabbit skeletal muscle 90% CS-TFC02 1 x 1 mg AAN02-S 1 x 125 µl Product Cat. # Amount (Ca2+ sensitive complex) 7A8.2.1) polymerization. Its versatility allows the study of the Ebashi Complex 0 Actin Polymerization 30-100 Anti-Pan Actin Mouse Monoclonal Antibody (Clone effects on polymerization (or depolymerization) of 0 5 10 15 20 25 30 BK003 (complex of tromyosin/ Bovine cardiac muscle 70% CS-TT05 1 x 1 mg AAN02 1 x 500 µl Assay Biochem Kit™ assays tropomodulin ) 7A8.2.1) a compound, tissue extract, or protein of interest. Time (min)